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Comparing Antioxidant Capacity and Neuroprotective effects of Phaleria Macrocarpa Fruit extracts and Tocotrienol Rich Fraction in Human Neuroblastoma Cells
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| Author:
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ASIAH BUDIMAN, NORFAIZATUL SHALIDA OMAR, WAN ROSALINA WAN ROSLI, GOON JO AAN
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| Abstract:
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Neurodegeneration occurs due to gradual loss of neuronal function and structure including neuron cell death associated with increased oxidative stress. Phenolic and flavonoid compounds in Phaleria macrocarpa fruit has been reported to exert antioxidant and anti-inflammatory activities which may intervene neurodegeneration. This study was conducted to compare the antioxidant capacity and neuroprotective effects of Phaleria macrocarpa ethanolic fruit extracts against tocotrienol rich fraction (TRF), which is a well-established antioxidant, in human neuroblastoma SH-SY5Y cells. Antioxidant properties of P. macrocarpa extracts and TRF were measured at different concentration using DPPH (2, 2-diphenyl-1-picrylhydrazil) assay. Cytotoxicity and neuroprotective properties of P. macrocarpa extracts and TRF were determined by measuring cell viability using MTS (3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulphenyl)-2H-tetrazolium) assay. The rate of apoptosis was measured using FITC Annexin V apoptosis by flow cytometer. Caspase-Glo assays were conducted to measure the activity of caspase-3, caspase-8 and caspase-9 which are mainly involved in the apoptotic pathway. Results showed that TRF has higher radical scavenging activity followed by BHT (DPPH standard) and then the P. macrocarpa fruit extracts. The MTS results demonstrated that P. macrocarpa fruit extracts (< 25 µg/ml) and TRF (= 15 µg/ml) were not toxic to neuron cells at low concentrations. Post-treatment of P. macrocarpa extracts (5 µg/ml) or TRF (1 and 3 µg/ml) exerted significant protection against BSO-induced cell death shown in cell viability studies. Post-treatment of P. macrocarpa fruit extracts also significantly reduced caspase-8 and caspase-9 activity but not caspase-3 activity against BSO-induced cell death. The findings suggest that P. macrocarpa extracts exert free radical scavenging activity similar to TRF and protected neurons BSO-induced cell death by inhibiting caspase-8 and caspase-9.
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| Keyword:
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Phaleria Macrocarpa, Antioxidant, Cytotoxic Activity, Neurodegeneration, Caspase.
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| EOI:
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| DOI:
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https://doi.org/10.31838/ijpr/2020.12.03.291
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| Download:
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